Enumeration of lymphocyte subsets using flow cytometry: Effect of storage before and after staining in a developing country setting |
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| Authors: | Sanju Jalla Sunil Sazawal Salkat Deb robert E Black Satya Narayan Das Archana Sarkar Maharaj K Bhan |
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| Institution: | (1) Division of Oncology and Hematology, Department of Medicine, Rush University Medical Center, 1653 West Congress Parkway, Chicago, IL 60612, USA |
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| Abstract: | Lymphocyte subset estimations by flow cytometry in population-based studies require transportation of samples from the field
site to the laboratory. As samples arrive late in the day they have to wait overnight before being processed. The effect of
two possible approaches, sample storage for 24 h before staining and immediate staining with analysis after 24 h and 48 h
were evaluated. Two sets of experiments were performed with EDTA (ethylenediamine tetra-acetate) anticoagulated peripheral
blood. In the first experiment, after collection, each sample was divided into two portions. One portion was stained at the
time of blood collection and the other 24 h later after keeping it at room temperature (38–45°C). In the second experiment,
blood samples were stained within 1–2 h. Each sample was analyzed immediately upon completion of staining process and subsequently
after 24 h and 48 h of storage at 4°C. Results suggest that blood collected in EDTA can be processed using whole blood lysis
method, after storage at room temperature (38–45°C) for 24 h with some but not significant alteration in T-cell subsets. Storage
at 4°C after staining for 24 h results in a lesser and insignificant loss of cells or alteration of T-cell subsets and may
be the method of choice. |
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| Keywords: | Lymphocyte storage flow cytometry leukocyte phenotypes |
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